LOCUS       sequence-340754-        4404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRMCD77 with S. flexneri 2a wzzBSF gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4404)
  AUTHORS   Daniels C, Morona R
  TITLE     Analysis of Shigella flexneri wzz (Rol) function by mutagenesis and 
            cross-linking: wzz is able to oligomerize.
  JOURNAL   Mol Microbiol. 1999 Oct;34(1):181-94. doi: 
            10.1046/j.1365-2958.1999.01591.x.
  PUBMED    10540296
REFERENCE   2  (bases 1 to 4404)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1046/j.1365-2958.1999.01591"; journalName: "Mol Microbiol"; 
            date: "1999-10"; volume: "34"; issue: "1"; pages: "181-94"
COMMENT     Sequence Label: pRMCD78
FEATURES             Location/Qualifiers
     source          1..4404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     314..333
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        314..332
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             364..386
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1671..1689)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1685..1732
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1777..1795)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1863..1967
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1968..2036,2037..2828)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2186..2205)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2999..3587
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3488..3507
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3741..3758
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3773..3912
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3838..3857)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3998..4020
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4014..4205)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg
       61 gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt
      121 catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt
      181 atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac
      241 gacaggagca cgatcatgcg cacccgtggc caggacccaa cgctgcccga gatcgatctc
      301 gatcccgcga aattaatacg actcactata gggagaccac aacggtttcc ctctagaaat
      361 aattttgttt aactttaaga aggagatata catatgatga gagtagaaaa taataatgtt
      421 tctgggcaaa accatgaccc ggaacagatt gatttgattg atttactagt gcagttgtgg
      481 cgtggcaaga tgacaattat catttccgtc attgtggcta ttgccctggc tattggttat
      541 ttggcagtag cgaaggagaa atggacgtca acagcaatta tcactcagcc cgacgtgggg
      601 caaattgctg gctataacaa tgccatgaat gttatctatg gtcaggctgc accgaaagta
      661 tcggatttgc aggagacgtt aattggtcgc ttcagttctg ccttctctgc attagcagaa
      721 acgctggata atcaggaaga gccagaaaaa cttaccatcg aaccttctgt taagaaccag
      781 caattaccat tgactgtttc ttatgttggg caaactgcag agggcgcaca aatgaagttg
      841 gcccaataca ttcagcaagt tgatgataaa gtgaatcaag agctagaaaa ggatctcaag
      901 gacaacattg ctctgggacg gaaaaacttg caggactctt taagaaccca ggaagtggtc
      961 gcgcaggagc agaaagatct gcgtatccgt cagattcagg aagcgttgca gtatgcgaat
     1021 caggcgcagg tgacaaagcc acaggttcag cagactgaag atgtgacgca agatacgttg
     1081 ttccttctag ggagcgaagc gctggagtcg atgattaagc atgaagcgac tcgtccgttg
     1141 gtgttctcac caaactacta tcagacacgt caaaacctgt tggatattga aaaattaaag
     1201 tttgatgatc ttgatattca tgcttaccgc tatgtgatga aaccgacgtt acctattcgt
     1261 cgcgatagtc cgaaaaaggc aatcaccttg attctggcag tgcttctggg cggcatggtt
     1321 ggcgcgggga ttgtgttggg gcgtaacgct ctgcgtaatt acaacgcgaa gtaatattat
     1381 tgtgcattta agagaaacgg gcagggtggt gacaccatgc ccgttttttt tgccggatgc
     1441 gatgctggcg catcttatcc ggcctacgtg tgttgagata atgtgtaggc acgataagtt
     1501 tgcgcatcgg gcaatggctc cgggtgtgac aacaacatca cacctgctcg gatccactag
     1561 taacggccgc cagtgtgctg gaattctgca gatatccatc acactggcgg ccgctcgagc
     1621 agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     1681 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     1741 aggaactata tccggataat tcttgaagac gaaagggcct cgtgatacgc ctatttttat
     1801 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     1861 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     1921 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     1981 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     2041 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     2101 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     2161 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     2221 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     2281 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     2341 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     2401 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     2461 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
     2521 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     2581 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     2641 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     2701 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     2761 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     2821 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     2881 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     2941 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3001 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3061 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3121 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     3181 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3241 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3301 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3361 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3421 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3481 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3541 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3601 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3661 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     3721 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     3781 ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac
     3841 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     3901 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     3961 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     4021 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     4081 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     4141 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     4201 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
     4261 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
     4321 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
     4381 caatgccagc gcttcgttaa taca
//
